Purification of a sixth ferredoxin from <i>Rhodobacter capsulatus</i>

Naud, Isabelle; Vincon, Mathilde; Garin, Jérôme; Gaillard, Jacques; Forest, Éric; Jouanneau, Yves

doi:10.1111/j.1432-1033.1994.tb18942.x

Cited by 33 publications

(27 citation statements)

References 35 publications

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“…This variation makes it impossible to use biochemical similarity to resolve bacterial phylogeny (62). Further, it does not appear likely that the E. histolytica, G. lamblia, and T. vaginalis FD came from the same bacterium, unless that bacterium had multiple FD genes as has been described for Rhodobacter capsulatus (six) (42). Nor does it appear likely that the E. histolytica POR, ADHE, and PPFK genes came from the same bacterium as those of G. lamblia, unless the bacterium had multiple POR, ADHE, and PPFK genes, and different fermentation genes were lost from E. histolytica and G. lamblia (3,26,48).…”

Section: Discussionmentioning

confidence: 99%

Evidence for the bacterial origin of genes encoding fermentation enzymes of the amitochondriate protozoan parasite Entamoeba histolytica

Rosenthal¹,

Mai²,

Caplivski³

et al. 1997

J Bacteriol

103

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Entamoeba histolytica is an amitochondriate protozoan parasite with numerous bacterium-like fermentation enzymes including the pyruvate:ferredoxin oxidoreductase (POR), ferredoxin (FD), and alcohol dehydrogenase E (ADHE). The goal of this study was to determine whether the genes encoding these cytosolic E. histolytica fermentation enzymes might derive from a bacterium by horizontal transfer, as has previously been suggested for E. histolytica genes encoding heat shock protein 60, nicotinamide nucleotide transhydrogenase, and superoxide dismutase. In this study, the E. histolytica por gene and the adhE gene of a second amitochondriate protozoan parasite, Giardia lamblia, were sequenced, and their phylogenetic positions were estimated in relation to POR, ADHE, and FD cloned from eukaryotic and eubacterial organisms. The E. histolytica por gene encodes a 1,620-amino-acid peptide that contained conserved iron-sulfur-and thiamine pyrophosphate-binding sites. The predicted E. histolytica POR showed fewer positional identities to the POR of G. lamblia (34%) than to the POR of the enterobacterium Klebsiella pneumoniae (49%), the cyanobacterium Anabaena sp. (44%), and the protozoan Trichomonas vaginalis (46%), which targets its POR to anaerobic organelles called hydrogenosomes. Maximum-likelihood, neighbor-joining, and parsimony analyses also suggested as less likely E. histolytica POR sharing more recent common ancestry with G. lamblia POR than with POR of bacteria and the T. vaginalis hydrogenosome. The G. lamblia adhE encodes an 888-amino-acid fusion peptide with an aldehyde dehydrogenase at its amino half and an iron-dependent (class 3) ADH at its carboxy half. The predicted G. lamblia ADHE showed extensive positional identities to ADHE of Escherichia coli (49%), Clostridium acetobutylicum (44%), and E. histolytica (43%) and lesser identities to the class 3 ADH of eubacteria and yeast (19 to 36%). Phylogenetic analyses inferred a closer relationship of the E. histolytica ADHE to bacterial ADHE than to the G. lamblia ADHE. The 6-kDa FD of E. histolytica and G. lamblia were most similar to those of the archaebacterium Methanosarcina barkeri and the ␦-purple bacterium Desulfovibrio desulfuricans, respectively, while the 12-kDa FD of the T. vaginalis hydrogenosome was most similar to the 12-kDa FD of ␥-purple bacterium Pseudomonas putida. E. histolytica genes (and probably G. lamblia genes) encoding fermentation enzymes therefore likely derive from bacteria by horizontal transfer, although it is not clear from which bacteria these amebic genes derive. These are the first nonorganellar fermentation enzymes of eukaryotes implicated to have derived from bacteria.The endosymbiont hypothesis or serial endosymbiosis theory, one of the most exciting ideas in evolutionary biology, postulates that the mitochondria and chloroplasts of eukaryotic cells derive from an endocytosed ␣-purple bacterium and cyanobacterium, respectively ( Fig. 1) (18,19,35,63). Although bacterial rRNA genes as well as those encoding some structural protei...

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Section: Discussionmentioning

confidence: 99%

Evidence for the bacterial origin of genes encoding fermentation enzymes of the amitochondriate protozoan parasite Entamoeba histolytica

Rosenthal¹,

Mai²,

Caplivski³

et al. 1997

J Bacteriol

103

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“…Mutant FdI proteins were expressed in E. coli and anaerobically purified on DEAE-cellulose as previously described (Grabau et al, 1991). Further purification of FdI variants involved gel filtration chromatography (Schatt et al, 1989), hydrophobic chromatography (phenyl-Sepharose, Pharmacia) and ion-exchange FPLC (Naud et al, 1994).…”

Section: Methodsmentioning

confidence: 99%

Identification of Residues of Rhodobacter capsulatus Ferredoxin I Important for Its Interaction with Nitrogenase

Naud

Meyer

David

et al. 1996

European Journal of Biochemistry

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In Rhodobacter capsulatus, ferredoxin I (FdI) serves as natural electron donor to nitrogenase. In order to probe amino acid residues possibly involved in the interaction with dinitrogenase reductase, FdI was subjected to site-specific mutagenesis. A three-dimensional structure of Fdl was designed by computer modelling and used for selecting target residues. Mutant ferredoxins bearing substitutions of surface residues, as well as a variant having a Met2-Tyr replacement in the vicinity of one cluster, have been constructed. All FdI variants were expressed to similar levels both in Escherichia coli and in a FdIdeleted mutant of the natural host. Once purified, the mutant ferredoxins exhibited molecular and spectroscopic properties almost identical to wild-type FdI. Determination of the reduction potential of FdI by cyclic voltammetry gave an EA of -510 mV (pH 7.6) for both clusters, which is one of the lowest values reported for a 2[4Fe-4S] ferredoxin. Only the [Tyr2]FdI variant showed a significant difference in redox potential (AE', = -15 mV). Based on in vitro assays, a [Glu27, Glu281FdI double mutant exhibited a twofold decrease in the electron transfer rate to dinitrogenase reductase while the affinity of this mutant for the enzyme was barely affected. On the other hand, an Asp36-.His substitution resulted in a sevenfold increase of the apparent K,,, for dinitrogenase reductase. Unlike FdI and the other mutant ferredoxins, the [His36]FdI variant also failed to form a cross-linked complex with dinitrogenase reductase upon incubation with a carbodiimide. It is concluded that Asp36 in FdI probably participates in the interaction between the two protein partners. Nevertheless, all the FdI mutants proved competent in restoring a wild-type phenotype when expressed in a FdI-deleted mutant background, indicating that none of the studied residues was absolutely critical for electron transfer to nitrogenase.

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“…Therefore, we named this C. crescentus ferredoxin gene fdxP. More recently, a third [2Fe-2S] ferredoxin has been discovered in Rhodobacter capsulatus that was shown to be 60% identical to the C. crescentus FdxP protein (16).…”

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confidence: 99%

A ferredoxin, designated FdxP, stimulates p-hydroxybenzoate hydroxylase activity in Caulobacter crescentus

1995

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A gene, fdxP, was identified upstream of the rrnA gene in Caulobacter crescentus and shown to encode ferredoxin II (FdII) by insertional inactivation. FdII is homologous to a class of [2Fe-2S] ferredoxins typified by putidaredoxin. Furthermore, reconstitution assays demonstrated that FdII was able to promote p-hydroxybenzoate hydroxylase activity in ferredoxin-depleted extracts. Thus, biodegradation of p-hydroxybenzoate may be ferredoxin dependent in C. crescentus.

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Purification of a sixth ferredoxin from Rhodobacter capsulatus

Cited by 33 publications

References 35 publications

Evidence for the bacterial origin of genes encoding fermentation enzymes of the amitochondriate protozoan parasite Entamoeba histolytica

Evidence for the bacterial origin of genes encoding fermentation enzymes of the amitochondriate protozoan parasite Entamoeba histolytica

Identification of Residues of Rhodobacter capsulatus Ferredoxin I Important for Its Interaction with Nitrogenase

A ferredoxin, designated FdxP, stimulates p-hydroxybenzoate hydroxylase activity in Caulobacter crescentus

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